中文摘要：

为了进一步从蛋白水平上检测Ndfip1（Nedd4 family interacting protein 1）和Nedd4（Neuronal precursor cell-expressed developmentally down-regulated4）在肢带型肌营养不良（Limb．girdle muscular dystrophy，LGMD）患者的肌肉组织中的表达及功能。以正常人的脑和肾脏组织的总RNA为模板．RT-PCR扩增Ndfip1和Mdd4部分编码序列。构建原核表达重组质粒pET28b-hNdfip1和pET28b—hNedd4，转化大肠杆菌BL21后．加IPTG诱导蛋白表达．Ni-NTA层析柱纯化蛋白，免疫BALB／c小鼠制备抗体．Western blot检测显示抗体能识别人和小鼠的Ndfip1和Nedd4，小鼠各组织中Ndfip1的蛋白表达谱与文献报道的Ndfip1 mRNA表达谱不一致，暗示Ndfip1具有转录和翻译水平的调控．

英文摘要：

To further verify Ndfipl and Nedd4 expression at protein level in the muscle of Limb-girdle muscular dystrophy （LGMD） patient and the function of these two genes, the partial coding sequences of Ndfipl and Nedd4 was obtained by RT-PCR, then recombined into prokaryotic expression vector pET28b and transformed into E.coli BL21 for expression. The purified proteins were gained by Ni affinity chromatography. The BALB/c mice were immunized with these purified proteins. The antibodies were successfully prepared and they can recognize human and mouse homologous protein. Western blotting of Ndfipl doesn＇t Ndfipl undergo in accord with Northern blotting result that reported in other paper,which suggested that regulation at both transcriptional and translational level.